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Papers In Press, published online ahead of print October 29, 2001
Massey Cancer Center, Virginia Commonwealth University, Richmond, Virginia 23298-0037
Corresponding Author: gdginder{at}hsc.vcu.edu
The methylation pattern of a 248-bp proximal transcribed region (rho248) of the avian embryonic rho-globin gene was found to correlate inversely with stage-specific expression in avian erythroid cells. In vitro methylation of the rho248 segment alone (in the absence of promoter methylation) resulted in a 5-fold inhibition of transcription in a transient transfection assay in primary erythroid cells in which the transfected gene is packaged into nucleosomal chromatin. This effect was observed if the rho248 segment was positioned adjacent to the promoter, but not when it was located 2.7 kb downstream. Fully methylated but not unmethylated rho248 formed a novel cell type-specific Methyl Cytosine binding Protein Complex (MeCPC) which contained Methyl Binding Domain protein-2 (MBD-2) and HDAC1 proteins but differs from MeCP-1. The histone deacetylase inhibitor Trichostatin A (TSA) failed to relieve methylation mediated repression of transcription from the rho-gene promoter, supporting the notion of the dominance of methylation over histone deacetylation in silencing through CpG rich sequences at this locus. These data demonstrate that site-specific methylation of a vertebrate gene 5' transcribed region alone at the exact CpG's that are methylated in vivo can suppress transcription in homologous primary cells and facilitate binding to a cell-type specific MeCPC.
J. Biol. Chem, 10.1074/jbc.M105580200
Submitted on June 18, 2001
Revised on October 29, 2001
Accepted on October 29, 2001
Methylation of promoter proximal transcribed sequences of an embryonic globin gene inhibits transcription in primary erythroid cells and promotes formation of a cell type-specific methyl cytosine binding complex
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